Regional hydrodynamic gene delivery to the rat liver with physiological volumes of DNA solution
Article first published online: 26 APR 2004
Copyright © 2004 John Wiley & Sons, Ltd.
The Journal of Gene Medicine
Volume 6, Issue 6, pages 693–703, June 2004
How to Cite
Zhang, X., Dong, X., Sawyer, G. J., Collins, L. and Fabre, J. W. (2004), Regional hydrodynamic gene delivery to the rat liver with physiological volumes of DNA solution. J. Gene Med., 6: 693–703. doi: 10.1002/jgm.595
- Issue published online: 25 MAY 2004
- Article first published online: 26 APR 2004
- Manuscript Accepted: 18 MAR 2004
- Manuscript Revised: 13 FEB 2004
- Manuscript Received: 24 NOV 2003
- Biotechnology and Biological Sciences Research Council
- Wellcome Trust
- gene therapy;
- portal vein;
The major barrier to the clinical application of hydrodynamic gene delivery to the liver is the large volume of fluid required using standard protocols. Regional hydrodynamic gene delivery via branches of the portal vein has not previously been reported, and we have evaluated this approach in a rat model.
The pGL3 plasmid with the luciferase reporter gene was used at 50 µg/ml in isotonic solutions, and was administered with a syringe pump for precise control of the hydrodynamic conditions evaluated. Gene expression was individually measured in six anatomically distinct liver lobes. The effect of systemic chloroquine to promote endocytic escape and a (Lys)16-containing peptide to condense the DNA into ∼100-nm nanoparticles was also evaluated.
Hydrodynamic conditions for excellent gene delivery were obtained by using 3-ml volumes (∼12 ml/kg) of isotonic DNA solution delivered at 24 ml/min to the right lateral lobe (∼20% of the liver mass). Under these conditions, >95% of gene delivery usually occurred in the targeted right lateral lobe. Outflow obstruction was essential for gene delivery, both at optimal and at very low levels of hydrodynamic gene delivery. The use of systemic chloroquine to promote endocytic escape did not augment hydrodynamic gene delivery, while condensation of DNA in non-ionic isotonic solutions (5% dextrose) to nanoparticles of ∼100 nm completely abolished gene delivery.
Regional hydrodynamic gene delivery via a branch of the portal vein offers a physiological model of liver gene therapy, for experimental and clinical application. Copyright © 2004 John Wiley & Sons, Ltd.